DMT1-Mutant Erythrocytes have Shortened Life Span, Accelerated Glycolysis and Increased Oxidative Stress

• Published in: Cellular physiology and biochemistry Vol. 34; no. 6; pp. 2221 - 2231
• Main Authors: Zidova, Zuzana, Kapralova, Katarina, Koralkova, Pavla, Mojzikova, Renata, Dolezal, Dalibor, Divoky, Vladimir, Horvathova, Monika
• Format: Journal Article
• Language: English
• Published: Basel, Switzerland Cell Physiol Biochem Press GmbH & Co KG 01.01.2014
• Subjects: Adenosine Diphosphate - blood; Adenosine Triphosphate - blood; Anemia, Hypochromic - blood; Anemia, Hypochromic - genetics; Anemia, Hypochromic - pathology; Animals; Apoptosis - genetics; Cation Transport Proteins - blood; Cation Transport Proteins - deficiency; Cation Transport Proteins - genetics; Disease Models, Animal; DMT1; Eryptosis; Erythrocytes - metabolism; Erythrocytes - pathology; Erythropoiesis; Glycolysis; Humans; Mice; Mutation; Original Paper; Oxidative Stress; Reactive Oxygen Species - blood; Eryptosis; Oxidative stress; DMT1
• ISSN: 1015-8987; 1421-9778
• DOI: 10.1159/000369665

Background/Aims: Deficiency of the divalent metal transporter 1 (DMT1) leads to hypochromic microcytic anemia. We have previously shown that DMT1 deficiency impairs erythroid differentiation and induces apoptosis of erythroid cells. Here we analyzed metabolic processes and survival of mature erythrocytes in order to address potential involvement of erythrocyte defect in the pathophysiology of the disease. Methods: FACS analysis was used to determine the half-life of erythrocytes (CFSE fluorescence), phosphatidylserine exposure (Annexin V binding), cytosolic Ca 2+ (Fluo3/AM fluorescence) and reactive oxygen species (ROS; DCF fluorescence). Enzyme activities were determined by standard biochemical methods. The concentration of ATP and ADP was measured on HPLC-MS/MS. Results: We observed an accelerated clearance of CFSE-labeled DMT1-mutant erythrocytes from circulating blood when compared to wild-type erythrocytes. In vitro, DMT1-mutant erythrocytes showed significantly increased Annexin V binding after exposure to hyperosmotic shock and glucose depletion. Despite exaggerated anti-oxidative defense, higher ROS levels were present in DMT1-mutant erythrocytes. Accelerated anaerobic glycolysis and reduced ATP/ADP ratio detected in DMT1-mutant erythrocytes indicate enhanced demand for ATP. Conclusions: We propose that DMT1 deficiency negatively affects metabolism and life span of mature erythrocytes; two other aspects of defective erythropoiesis which contribute to the pathophysiology of the disease.